幽门螺杆菌黏附素基因babA_2的克隆表达及定位

Cloning high expression and location of H.pylori adhesion babA_2 gene

目的 :克隆并表达H .pylori黏附素babA2 基因 .方法 :提取H .pylori染色体基因 ,用PCR方法扩增babA2 基因 ,将其克隆至表达载体pET 2 2b(+) ,并在BL2 1 (DE3)大肠杆菌中表达及定位分析 .结果 :分离得到了 2 .2kb的babA2基因片段 ,在 37℃诱导表达 3h后 ,重组蛋白表达量占菌体总蛋白的 34.8% ,其中分泌表达占周质总蛋白的 2 2 .7% ,可溶性表达占上清的 1 5 .0 % ,包涵体占沉淀的 86 .7% .结论 :H .py loribabA2 基因的克隆与表达为H .

AIM: To isolate and express bab A 2 gene from H.pylori. METHODS: bab A 2 gene was amplified from H.pylori chromosome by PCR and inserted into the prokaryotic expression vector pET 22b(+).The recombinant plasmid was transformed into the BL21(DE3) E.coli strain. RESULTS: The 2.2 kb bab A 2 gene was successfully isolated. Recombinant E.coli strains expressed bab A 2 were obtained, the expression protein amounted to 34.8% of the total bacterial protein , 22.7% of the bacterial periplasm protein , 15.0% of the bacterial sonicate supernatant and 86.7% of the bacterial inclusion body after being induced with IPTG for 3 h at 37℃. CONCLUSION: Cloning and high expression of bab A 2 gene lay the foundation for further research in the molecular mechanism of H.pylori adhesin and constructing the H.pylori vaccine.